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Overexpression of AtYUCCA6 in soybean crop results in reduced ROS production and increased drought tolerance

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Abstract

Drought is a major abiotic stress in crop yield and its inevitable consequence is the increased production of reactive oxygen species (ROS) and cell damage. To reduce excessive ROS accumulation in soybean, AtYUCCA6 gene was transformed via Agrobacterium-mediated transformation. About 3% of transformation efficiency was generated from five batches of the transformation experiment. Eighteen transgenic plants were produced with PPT resistance and analyzed for introgression of AtYUCCA6. T-DNA insertion and expression were confirmed by PCR, Southern blot and reverse transcriptase-PCR. In the drought tolerance tests with transgenic lines #2, #3, and #5, all three lines were less affected by drought treatment and survived in the water-deficit conditions while non-transgenic plants did not survive under the same drought condition. The physiological aspects of transgenic lines were also much stronger than NT plants by showing higher chlorophyll content and lower ion leakage during water-deficit conditions (p < 0.01), indicating the prevention of cell-membrane damage. Measurement of transpiration rate on detached leaves from transgenic plants showed nearly 10% less water loss. Finally, 3 transgenic lines (#2, #3, and #5) were investigated for ROS accumulation by DAB staining of detached leaves under water-deficit conditions. Unlikely NT plants with severe dark browning after 14 days of drought treatment, transgenic lines #2, #3, and #5 did not show significant browning.

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Acknowledgements

This work was supported by the Next-Generation BioGreen 21 Program, Rural Development Administration (PJ01366501 granted to Y. S. Chung), and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2017R1A6A3A11028883 granted to H. J. Kim) in the Republic of Korea.

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  1. Jin Sol Park and Hye Jeong Kim contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Genetics, College of Natural Resources and Life Science, Dong-A University, 37, Nakdong-daero 550beon-gil, Saha-gu, Busan, 49315, Republic of Korea

    Jin Sol Park, Hye Jeong Kim, Hyun Suk Cho, Ho Won Jung & Young-Soo Chung

  2. Division of Applied Life Science (BK21Plus), Plant Molecular Biology and Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju, 52828, Republic of Korea

    Joon-Young Cha

  3. Department of Biomedical Science and Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 05029, Republic of Korea

    Dae-Jin Yun

  4. National Institute of Agricultural Science, Rural Development Administration, Jeonju-si, 54875, Republic of Korea

    Seon-Woo Oh

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  1. Jin Sol Park
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11816_2019_527_MOESM1_ESM.tif

Supplementary material 1 Fig. S1. Vector used for soybean transformation. Amplified AtYUCCA6 (1,281 bp size) was subcloned into pPZP-3’PinII-Bar vector for soybean transformation. LB/RB, left/right T-DNA border; p35S/T35S, CaMV (cauliflower mosaic virus) 35S promoter/terminator; 3’PinII/3’nos, terminator; Bar, coding region of the DL-phosphinothricin resistance gene. HindIII and PstI restriction enzyme sites are marked (TIF 63 KB)

11816_2019_527_MOESM2_ESM.tif

Supplementary material 2 Fig. S2. AtYUCCA6overexpression soybean transgenic plants. (a) Co-cultivation of half-seed explants after inoculation (left) and at 5 days after inoculation (right). (b) Shoot induction on SIM without PPT for 14 days. (c) Shoot induction on SIM with 10 mg l-1 PPT for another 14 days. (d) Shoot elongation on SEM with 5 mg l-1 PPT. (e) Root formation (f) Acclimation of putative transgenic plant in a small pot. (g) Transgenic plant (T0) grown in a large pot in the greenhouse. (h) Herbicide (100 mg l-1 PPT) painting showing sensitivity in NT plant leaves (left) and resistance in transgenic plant leaves (right) (TIF 1279 KB)

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Park, J.S., Kim, H.J., Cho, H.S. et al. Overexpression of AtYUCCA6 in soybean crop results in reduced ROS production and increased drought tolerance. Plant Biotechnol Rep 13, 161–168 (2019). https://doi.org/10.1007/s11816-019-00527-2

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